LOCUS       sequence-444960-        4795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RepA(N99D) SC101 T0-BS7 proD SD8 sfGFP( V93AGGA Y151UAGA) ProK 5 
            tRNA operon tetA-orfL bla.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4795)
  AUTHORS   Costello A, Peterson AA, Lanster DL, Li Z, Carver GD, Badran AH
  TITLE     Efficient genetic code expansion without host genome modifications.
  JOURNAL   Nat Biotechnol. 2024 Sep 11. doi: 10.1038/s41587-024-02385-y.
  PUBMED    39261591
REFERENCE   2  (bases 1 to 4795)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2024 Sep 11. doi: 10.1038/s41587-024-02385-y."
COMMENT     Sequence Label: pAMC042z21
FEATURES             Location/Qualifiers
     source          1..4795
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      998..1220
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2739..2756
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    3013..3029
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4693..4764)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttaaatacga ccgtatttcg ccaaaatgtt ttccaaggtg gtacgctgtt tctgggtcaa
       61 ccaagagaaa gaaccgttca aatcatgttt agactgcata tcagacaaga atttcgcttc
      121 gaaagaggtc aactgaattt tcgcggtcaa cgcatcatgc aaggttttac gcaaaccgtt
      181 acgcaagtaa gaatcagagg taatggtggt cggaatttta tcaccgttca ttttaatctg
      241 gttgttttcc aattcggtaa ccaaatccat ctgacgatcc aattcaacct ggaaaatcaa
      301 ggtatcggtc ggacgaccac gtttatcaac aaccaatttc atgttagagt aggtgttcaa
      361 atctttagaa atcggtttca aaacccactg gttcaaacgt ttgaattcat ggtagttgtt
      421 ttccaacatc aacatgaatt tgaattcatc caaagaaatt tcaatgttcg ctttatgggt
      481 ttttttctgg gtcaattctt tcaacaacca ttcgtaaata cgcatagagt atttgttttc
      541 gaaagattta acatgttcca agttgtattt aatgaatttt ttcaactgga acaagtacgg
      601 caaaatttct tcagagaaaa ccaattccaa tttttcagaa gagaatttcg cgtaatcggt
      661 ccactggaaa atttcgaaac ctttaaccaa cgggttacga atttcaacgg tacgggtcat
      721 caattcacgg gtcgctttcg ccaaaacacc gtacgcgttt tcacgagaaa tgttcatcat
      781 ctgcgcgtac tggttgtagg tgaaagaaac ggtacgttct ttacgggtcg ggttttcaat
      841 ggtcgggttc aacaacgcaa cacaacacaa aatcaatttg gtttcatgtt cggtcaaatc
      901 gtaacgagaa atcgccaatt cgttcgcttt gaaaacaacc aattcagaca tacatctcaa
      961 ttgtctaggt gattttaatc actataccaa ttgagatggg ctagtcaatg ataattacta
     1021 gtccttttcc tttgagttgt gggtatctgt aaattctgct agacctttgc tggaaaactt
     1081 gtaaattctg ctagaccctc tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat
     1141 attcaagtgg ttataattta tagaataaag aaagaataaa aaaagataaa aagaatagat
     1201 cccagccctg tgtataactc actactttag tcagttccgc agtattacaa aaggatgtcg
     1261 caaacgctgt ttgctcctct acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc
     1321 ctcgcaagct cgggcaaatc gctgaatatt ccttttgtct ccgaccatca ggcacctgag
     1381 tcgctgtctt tttcgtgaca ttcagttcgc tgcgctcacg gctctggcag tgaatggggg
     1441 taaatggcac tacaggcgcc ttttatggat tcatgcaagg aaactaccca taatacaaga
     1501 aaagcccgtc acgggcttct cagggcgttt tatggcgggt ctgctatgtg gtgctatctg
     1561 actttttgct gttcagcagt tcctgccctc tgattttcca gtctgaccac ttcggattat
     1621 cccgtgacag gtcattcaga ctggctaatg cacccagtaa ggcagcggta tcatcaacag
     1681 gcttacccgt cttactgtca agaggacatc cggtttgttc agaacgctcg gtcttgcaca
     1741 ccgggcgttt tttctttgtg agtccatagt acacttgaat aaataaaaac agccgttgcc
     1801 agaaagaggc acggctgttt ttattttaga cttagggacc ctcacagcta acaccacgtc
     1861 gtccctatct gctgccctag gtctatgagt ggttgctgga taactttacg ggcatgcata
     1921 aggctcgtat aatatattca gggagaccac aacggtttcc ctctacaaat aattttgttt
     1981 aactttacct gcaggtgcag taaggaggaa aaaaaaatgt ctaaaggtga agaattgttc
     2041 accggtgttg ttccgatttt ggttgaattg gatggtgatg ttaacggtca taaattctct
     2101 gttcgtggtg aaggtgaagg tgatgcgacc aacggtaaat tgaccttgaa attcatttgt
     2161 accaccggta aattgccggt tccgtggccg accttggtta ccaccttgac ctacggtgtt
     2221 cagtgtttct ctcgttaccc ggatcatatg aaacagcatg atttcttcaa atctgcgatg
     2281 ccggaaggtt acaggacagg aacgtaccat ttctttcaaa gatgatggta cctacaaaac
     2341 ccgtgcggaa gttaaattcg aaggtgatac cttggttaac cgtattgaat tgaaaggtat
     2401 tgatttcaaa gaagatggta acattttggg tcataaattg gaatacaact tcaactctca
     2461 taacgtttag aattaccgcg gataaacaga aaaacggtat taaagcgaac ttcaaaattc
     2521 gtcataacgt tgaagatggt tctgttcagt tggcggatca ttaccagcag aacaccccga
     2581 ttggtgatgg tccggttttg ttgccggata accattactt gtctacccag tctgcgttgt
     2641 ctaaagatcc gaacgaaaaa cgtgatcata tggttttgtt ggaattcgtt accgcggcgg
     2701 gtattaccca tggtatggat gaattgtaca aaggttctca tcatcatcat catcattaaa
     2761 cttaattaac ggcactcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     2821 tgctgaggag acttagggac ccttgtgctt ctcaaatgcc tgaggccagt ttgctcaggc
     2881 tctccccgtg gaggtaataa ttgacgatat gatcagtgca cggctaacta agcggcctgc
     2941 tgactttctc gccgatcaaa aggcattttg ctattaaggg attgacgagg gcgtatctgc
     3001 gcagtaagat aattgtgagc ggataacaat tagcagacaa gataattttg cacccagcaa
     3061 acttggtacg taaacgcatc gtggcgaact ggtccgggac caccgagcat tcctaagctc
     3121 cggttagccg ggttcaactc ccgggttcgt cgccaaaatt tgaaagtgct gtaaggcaca
     3181 gaccacccaa gggggacggt ccggcgacca gcaggcctct ataagcctcg ccttgcgggg
     3241 ttcgacaccc cggtctctcg ccagtttaaa agacatcggc gtcaagcgga tgtctggctg
     3301 aaaggcctga agaatttgga cctgtgatca tgtagatcga gaggcttcta atccatatca
     3361 gccgggttag attcccggcg ggtccgccat ctcttacttg atatggcttt agtagcggta
     3421 tcaatatcag cagtaaaata aatttcccga tcccgcccta gctcagaggt agagcgtgcc
     3481 tctctagaag ggcatggtcc ccggttcaaa tcctgggggc gggaccaact actttatgta
     3541 gtctccgccg tgtagcaaga aattgagaag tgaactggtg gctcaatggt agagctgcac
     3601 gcttccgaac gtgttggttg caggttcaag tcctgtccag ttcaccaatt tattcaagac
     3661 gcttaccttg taagtgcacc cagtaggatc cgtcgacctg cagatcctta gcgaaagcta
     3721 aggatttttt ttaagcttct gccattcatc cgacttaatt aacggcactc ctcagcaaat
     3781 ataatgaccc tcttgataac ccaagagggc attttttaat gcccatggcg tttaccaatg
     3841 tttaatcaaa gacgcaccaa tttccgcaat ctgacggtta cgttcatcca tggtcgcctg
     3901 agaaccggtg gtgtaaataa caacaatacg agacggttta ccatccggac ccaacgccgc
     3961 aataatacca cgagaaccac gttcacccgc accagattta tccgcaatga accaacccgc
     4021 cggcaacgca gaacgcaaca acggacccgc aactttatcc gcttccatcc aatcaatcaa
     4081 ctgctgacga gacgccaagg tcaacaattc accggtcaac aatttacgca aggtggtcgc
     4141 catcgcaacc ggcatggtgg tatcacgttc atcgttcgga atcgcttcgt tcaattccgg
     4201 ttcccaacga tccaaacggg taacatgatc acccatgtta tgcaagaacg cggtcaattc
     4261 tttcggacca ccaatggtgg tcaacaacaa gttcgccgcg gtgttatcag acatggtaat
     4321 cgccgcagaa cacaattcac gaacggtcat accatcggtc aaatgttttt cggtaaccgg
     4381 agagtattca accaaatcgt tctgagagta atgaatacga cgacccaact gttcctgacc
     4441 cgcatcaata cgagacaaaa ccgcaccaca caacaaaact ttgaaggtag acatcatcgg
     4501 gaaacgttct tccggacgga aagattccaa aattttacca gagttcaaat ccaattcaat
     4561 gtaaccaaca cgcgcaccca actgatcttc cgcatcttta actttaacca aggtttccgg
     4621 atgcgcgaaa accggcaaac agaacgccgc gaagaacgga atcaacgcaa cacggaaatg
     4681 ctgaatagac atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4741 catgagcgga tacatatttg aatgtattta gaaaaataaa ggccaaataa gccgt
//
